Modulation of Cystatins and Cathepsins in HIV-1 Neuropathogenesis

半胱氨酸蛋白酶抑制剂和组织蛋白酶在 HIV-1 神经发病机制中的调节

• 批准号: 7493731
• 负责人: LOYDA M MELENDEZ
• 金额: $ 50万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7493731
• 关键词: Adenovirus Vector; Affect; Antibodies; Antiviral Therapy; Apoptotic; Behavioral; Biological Markers; Brain; Cathepsin C; Cathepsins; Cathepsins B; Cells; Cellular Structures; Cerebrospinal Fluid; Cognitive; Cystatins; Diagnosis; Disease; Effector Cell; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Freezing; Gene Expression; Gene Proteins; Goals; HIV; HIV Infections; HIV encephalitis; HIV-1; Highly Active Antiretroviral Therapy; Hispanics; Immune; Immunoprecipitation; Impaired cognition; Impairment; Infection; Inflammatory; Knowledge; Laboratories; Liquid substance; Maintenance; Measures; Microglia; Molecular; Molecular Profiling; Mononuclear; Motor; National NeuroAids Tissue Consortium; Nerve Degeneration; Neurocognitive; Neurons; Neuropathogenesis; Nitric Oxide; Nuclear; Oxidative Stress; Patients; Phagocytes; Phenotype; Plasma; Play; Predictive Value; Process; Production; Proteins; Proteomics; Reporting; Role; Sampling; Signal Pathway; Small Interfering RNA; Source; Testing; Up-Regulation; Viral; Viral Load result; Virus; Woman; antiretroviral therapy; brain tissue; cognitive function; cohort; in vivo; knock-down; macrophage; monocyte; neuropathology; neurotoxic; neurotoxicity; novel; post gamma-globulins; protein expression; research study; response; stefin

DESCRIPTION (provided by applicant): Modulation of cystatins and cathepsins in the neuropathogenesis of HIV-1 infection Cognitive impairment (CI) occurs during advanced HIV-1 infection, despite antiretroviral therapy (HAART). The molecular and cellular mechanisms for neuronal impairment evolve from neurotoxic secretory products produced from mononuclear phagocytes (MP; perivascular macrophages and microglia). Although the intracellular mechanisms that affect toxic MP secretions are incompletely known, the effector cell responses do play a pivotal role in CI. The long-term goal of the proposed study is to provide a better understanding of the role of macrophage secretory factors in homeostatic control during progressive HIV infection of the brain and to uncover novel targets for potential therapies. Among many secretory factors, cystatins and cathepsins secreted by MP play broad yet important roles in neuroregulatory responses. In particular, our laboratory and ex vivo experiments have shown the presence of cathepsins, cystatins, and SOD in virus-infected MP culture fluids and cerebrospinal fluid (CSF) of HIV-seropositive women with CI by proteomic analyses. Nevertheless, their role in neurodegenerative processes is not completely understood. The proposed study will elucidate the role played by cystatins and cathepsins, and SOD, on HIV-infected macrophages and their interactions with neuronal cells, which is critical for controlling the pro-inflammatory microenvironment in the brain during the induction of CI. We hypothesize that dysregulation of cystatins in HIV-infected macrophages during oxidative stress promotes cathepsin-B induced neurotoxicity and therefore contributes to CI. We will test this hypothesis with the following specific aims: 1) To determine, by analysis of gene and protein expression, how the interplay between cystatin B and cathepsin B regulates HIV-1 infection in macrophages and affects neurotoxic phenotype; 2) To define the roles of cystatin B/C, cathepsin B, SOD, and proteins affected by their deregulation, found in Aim 1, as predictors of progression to CI in the presence of HAART. The significance and long-term implication of this study is that it will fill the gaps in knowledge regarding the interactions between cystatins, cathepsins and SOD in macrophages and their role in neuropathogenesis. The studies with the Hispanic cohort will also clarify the role of cystatins, cathepsins, and SOD in HIV-cognitive impairment in the post-HAART era. The proposed studies will determine the role of macrophage cystatins, cathepsins, SOD, and the proteins affected by their deregulation in HIV replication and neurotoxicity. The Hispanic cohort of HIV-infected women characterized for cognitive function and under antiviral therapy is an excellent source for the study of these possible biomarkers in the HAART era. The CSF, plasma, monocyte, and macrophage samples derived from this cohort have been stored longitudinally and provide an important source for testing our hypothesis: that over expression of cystatins in HIV-infected macrophages will promote cathepsin B-induced neurotoxicity and therefore contribute to CI. This study may elucidate novel targets for diagnosis and therapy.

描述(由申请人提供)：半胱氨酸氨基转移酶和组织蛋白在HIV-1感染的神经发病机制中的调节作用尽管接受了抗逆转录病毒治疗(HAART)，但认知障碍(CI)在晚期HIV-1感染期间发生。神经元损伤的分子和细胞机制由单核巨噬细胞(MP；血管周围巨噬细胞和小胶质细胞)产生的神经毒性分泌产物进化而来。尽管影响毒性MP分泌的细胞内机制尚不完全清楚，但效应细胞反应在CI中确实起着关键作用。这项拟议研究的长期目标是更好地了解巨噬细胞分泌因子在进行性HIV脑感染期间体内平衡控制中的作用，并发现潜在治疗的新靶点。在众多的分泌因子中，MP分泌的半胱氨酸和组织蛋白在神经调节反应中发挥着广泛而重要的作用。特别是，我们的实验室和体外实验表明，通过蛋白质组学分析，在感染病毒的MP培养液和HIV血清阳性CI妇女的脑脊液(CSF)中存在组织蛋白酶、半胱氨酸氨基转移酶和超氧化物歧化酶。然而，它们在神经退化过程中的作用并不完全清楚。这项拟议的研究将阐明半胱氨酸蛋白酶和组织蛋白酶以及超氧化物歧化酶在HIV感染的巨噬细胞中所起的作用以及它们与神经细胞的相互作用，这对于控制脑梗塞诱导过程中的促炎微环境至关重要。我们假设，在氧化应激期间，HIV感染的巨噬细胞中半胱氨酸氨基转移酶的失调促进了组织蛋白酶-B诱导的神经毒性，从而促进了CI的发生。我们将通过以下具体目标来检验这一假说：1)通过基因和蛋白质表达的分析，确定胱抑素B和组织蛋白酶B之间的相互作用如何调节巨噬细胞中的HIV-1感染并影响神经毒性表型；2)确定在目标1中发现的胱抑素B/C、组织蛋白酶B、超氧化物歧化酶以及它们的放松调控所影响的蛋白质作为HAART存在时进展为CI的预测因子的作用。这项研究的意义和长期意义在于，它将填补有关巨噬细胞中半胱氨酸氨基转移酶、组织蛋白和超氧化物歧化酶之间相互作用及其在神经发病机制中作用的知识空白。与西班牙裔队列的研究还将阐明半胱氨酸氨基转移酶、组织蛋白和超氧化物歧化酶在后HAART时代艾滋病毒认知障碍中的作用。这项拟议的研究将确定巨噬细胞半胱氨酸氨基转移酶、组织蛋白酶、超氧化物歧化酶及其解除调控所影响的蛋白质在艾滋病毒复制和神经毒性中的作用。在HAART时代，以认知功能为特征并接受抗病毒治疗的西班牙裔感染妇女队列是研究这些可能的生物标记物的极好来源。来自这个队列的脑脊液、血浆、单核细胞和巨噬细胞样本已经被纵向存储，并为检验我们的假设提供了一个重要的来源：在HIV感染的巨噬细胞中过度表达胱抑素将促进组织蛋白酶B诱导的神经毒性，从而促进CI的发生。这项研究可能会阐明诊断和治疗的新靶点。